Often times in the scientific literature you will find that a topic suddenly becomes trendy. This might follow a technological advancement that opens up a new subject area, a serendipitous co-discovery or after an important publication opens a Pandoras box of questions. Over the past few months the Arabidopsis research literature has seen a number of exciting papers that have tackled different aspects of the plants response to heat, otherwise known as thermosensing.

These papers focus on two aspects of this response, either in heat perception or at the convergence point for the downstream cascade of signals. At the level of perception back-to-back papers published at the end of 2016 showed that phytochromeB, already a well known light receptor, also acts as a temperature sensor [1,2]. At the phenotypic level this involves the regulation of growth, as phyB mutants that have a single point mutation and are stuck in the Pfr form have longer hypocotyls. At the level of gene expression the mutant phyB protein constitutively binds to the promotors of warm-responsive genes. Temperature does not cause changes to PhyB mRNA or protein indicating that the protein directly act as a thermosensor. One of these papers show that PHYB negatively regulates the temperature-dependent expression of the PIF4 transcription factor [2] so in phyB mutants the longer hypocotyl can be traced, at least in part, due to extra PIF4 expression. This regulation is relevant to three more recent papers in which regulation of PIF4 activity is a central and recurring theme.

In the first of these papers Scott Hayes and colleagues investigate the response stimulated by UV light, which is perceived by the UVR8 receptor [3]. They show that whereas PIF4 is stimulated by increasing temperature, this is off-set by the activity of UVR8 which, when subject to high levels of UV-B, reduces PIF4 expression. A possible in vivo explanation for this response is provided in an associated comment piece [4] that postulates that in direct sunlight (where there is plentiful UV-B) there is no need for a hypocotyl elongation. However at high temperature without UV-B, the plant may interpret that it is growing in the shade so it might require a burst of hypocotyl elongation, which is mediated via PIF4.

The final two papers are from the lab of Vinod Kumar and focus on the activity of the PIF4 protein. Firstly they show that two well-studied signaling modules acting via either HY5 or DET1 differently impact PIF4 activity. Whereas DET1 directly interacts with PIF4, HY5 binds to the PIF4 recognition motif on its known DNA targets. At high temperature the HY5 protein is removed by the activity of the E3 ligase COP1, freeing up PIF4 expression. A previous study has shown UVR8 can sequester the COP1 protein so in high UV-B the activity of COP1 may be removed thus allowing HY5 to compete with PIF4 for its binding sites. Although the recent ‘UV-focused study’ [3] showed that HY5 gene expression was increased by UV-B, the role of HY5 in their experiments was slightly confused. The subtle and overlapping roles of these multiple proteins makes it challenging to obtain a complete picture of each of their roles independently of each other, which in any case would be an unrealistic in vivo situation.

As growth increases at higher temperature so does the plants suspectibility to pathogens, processes that are linked via PIF4. pif4 mutants are more resistant to infection and natural variation of the PIF4 gene provides a range of circumstances whereby plants become unresponsive to temperature change yet have higher levels of immunity. These findings have great significance for future crop improvement strategies that are extremely relevant for our warming world.

As with the integration of white light, UV-B and temperature signals, the link between temperature and disease is finely modulated to ensure that the plant maximizes its environmental situation and is able to rapidly adapt to its current conditions (such as on a windy day when the levels of shade and UV-B are highly variable).

This recent set of papers show that the PIF4 protein plays a central role through each adaptive molecular decision.

Vinod Kumar kindly provides an audio overview of this recent set of papers. Also on the GARNet YouTube Channel.

Todays Arabidopsis Research Roundup includes some excellent examples of UK labs engaged in collaborative work with researchers from around the globe. However first up is a study solely from the John Innes Centre, led by Vinod Kumar, that investigates the role of PIF4 during the thermosensory response. Secondly David Evans (Oxford Brookes University) is a co-author on a French-led study that has looked into the role of LINC complexes during interphase heterochromatin patterning. Thirdly is the description of the new PhenoTiki imaging tool that has come from the lab of Sotirios Tsaftaris in Edinburgh. Work from Paul Dupree (University of Cambridge) features in the ARR for the second consecutive week, this time with a study looking at the sugar composition of seed mucilage. The penultimate study is from the lab of Renier van der Hoorn (Oxford University) who investigates the role of Cys proteases during senescence and finally is a study from Seth Davis (University of York) that looks at the link between the circadian clock and the plants energy sensing mechanisms.

Open Access
Vinod Kumar (John Innes Centre) leads this study that looks at the role of the PHYTOCHROME INTERACTING FACTOR 4 (PIF4) transcription factor during the thermosensory response and its effect on plant architecture. They looked at the natural variation of PIF4, demonstrating the role of different varients on the balance between growth and immunity to pathogens. Pertubing PIF4-mediated effects result in temperature-resilient disease resistance. This study links with a paper highlighted in last weeks ARR from Kerry Franklin and co-authors that presented the role of UVR8 on the control of PIF4 heat responsive effects. These studies further confirm the important role of PIF4 in plant development in response to environmental change and biotic challenges.

Vinod discusses this paper and a related manuscript from next weeks ARR. Also available on the GARNet YouTube channel.

This study is led by Christophe Tatout from Clermond-Ferrand and includes David Evans and Axel Poulet (Oxford Brookes University) as co-authors. The paper focuses on the role of the nuclear envelope-localised LInker of Nucleoskeleton and Cytoskeleton (LINC) complex on nuclear morphology and interphase chromatin localisation. This work is underpinned by the use of novel 3D imaging tools to define where in the nucleus the chromatin is localised in both wildtype and linc mutant plants. This allows the authors to show that the LINC complex is necessary for proper heterchromatin organisation at the nuclear periphery, which might have broad implications for gene expression and transcriptional silencing.

Open Access
This manuscript describes the PhenoTiki tool that is designed for the automated phenotyping of Arabidopsis rosettes, work which is led by Sofortios Tsaftaris (University of Edinburgh). PhenoTiki describes both the imaging software and also cheap-to-use off-the-shelf hardware that allows for facile imaging at reduced costs. The proof-of-concept study in the paper shows a comprehensive analysis from a range of parameters in 24 Arabidopsis rosettes from different genotypes. This data is compared favourably to more expensive methods of automated phenotyping so the authors hope PhenoTiki can be adopted as a low-cost method for image analysis. Full details can be found at http://phenotiki.com.

Open Access
Paul Dupree (University of Cambridge) is part of this global collaboration with colleagues from Australia, USA and Chile. The study investigates the intracellular movement of the plant cell polysaccharide pre-cursor UDP-glucuronic acid (UDP-GlcA). To identify genes involved in this process they cleverly screened mutants for alteration in seed mucilage, which has high level of other polysaccharides. This strategy identified UUAT1, which is a golgi-localised transporter of UDP-GlcA and UDP-galacturonic acid (UDP-GalA). Uuat1 mutants have altered sugar composition in both the seed coat mucilage and in other plant organs. These changes were also associated with an increase, by a currently unknown mechanism, of homogalacturonan methylation. Overall the authors show that UUAT1 is important for the correct distribution of cell wall polysaccahrides throughout growing embryo and will surely play important developmental roles in the function of the cell wall.

In this paper Renier van der Hoorn (University of Oxford) interacts with US, German and Australian colleagues to use the activity-based protein profiling (ABPP) technique to assess the activity of active enzymes during senescence. They show that in Arabidopsis leaves the expression of several Papain-like Cys Proteases (PLCPs) is elevated but the activity of many Vacuolar Processing Enzymes (VPEs) is decreased, even though their transcript level increases. The amount of senescence was assessed in plants with mutations in different members of these protease families and surprisingly did not find any difference when compared to wildtype plants. One exception was in plants containing a mutation in the AALP PLCP which showed a significant, albeit slight, descrease in the rate of senescence.

Seth Davies (University of York) leads this study that includes German, Mexican and Chinese collaborators and looks at the link between the circadian clock and plant metabolism. The energy sensing Snf1 (sucrose non-fermenting 1)-related kinase 1 (SnRK1) complex contains the catalytic AKIN10 protein, which plays an important role in clock function by controlling expression of the key evening element GIGANTEA (GI). This AKIN10 effect requires the clock regulator TIME FOR COFFEE (TIC) demonstrating an important role for the plants energy sensing mechanisms, via the AKIN10, in conditional control of clock gene expression.

The first Arabidopsis Research Roundup of 2017 includes a wide range of studies that use our favourite model organism.

Firstly Kerry Franklin (University of Bristol) is the corresponding author on a paper that describes the complex interaction between the responses to sunlight and heat. Secondly Paul Dupree (University of Cambridge) leads a study that defines the important structural relationship between xylan and cellulose. Thirdly members of Gos Micklem’s group in Cambridge are part of the Araport team that present their ThaleMine tool.

Richard Napier (University of Warwick) is a co-author on the fourth paper that introduces a new chemical tool for study of the auxin response. The penultimate paper includes Matthew Terry (University of Southampton) on a paper that investigates the role of a Fe-S-containing protein cluster in chlorophyll biosynthesis and finally there is a methods paper from Stefanie Rosa in Caroline Dean’s lab at the John Innes Centre that describes the use of FISH to detect single molecules of RNA.

This collaboration between the research groups of Kerry Franklin (University of Bristol) and Gareth Jenkins (University of Glasgow) looks at how the perception of UV-B light inhibits the morphological changes that occur in response to increased temperatures (thermomorphogenesis). This response includes induced hypocotyl elongation, which is mediated via PIF4 and various players in the auxin response. Interestingly the authors show that UV-B light perceived by UVR8 attenautes this response by preventing PIF4 abundance and by stabilising the the bHLH protein LONG HYPOCOTYL IN FAR RED (HFR1) protein. These results suggest that there exists a precise mechanism for fine-tuning the growth responses that occur in sunlight that would usually include both increased temperature and UV-B irradiation.

http:/​/​dx.​doi.​org/10.1038/ncomms13902 Open Access
In this paper Paul Dupree (University Cambridge) collaborates both with colleagues in Spain and with his father Ray, who is a physicist at the University of Warwick. They use NMR to perform a structural analysis of xylan, which is the most prevalent non-cellulosic polysaccharide in the cell wall matrix and binds to cellulose microfibrils. Whereas in solution xylan forms a threefold helical screw, it flattens into a twofold helical screw ribbon to closely bind to cellulose when in the cell wall. They used the cellulose-deficient Arabidopsis irx3 mutant to show that the xylan two-fold screw confirmation breaks down when it cannot bind cellulose. The authors state that this finding has important implications in our understanding of the formation of the cell wall and perhaps more importantly how it might be broken down during attempts to maximise economic usages of plant biomass.

This paper is presented by the Araport team, which is based in the USA but includes representatives from Gos Micklem’s lab in University of Cambridge. They outline the functionality of the ThaleMine data warehouse which is an important component of the tools included on Araport (https://www.araport.org/). ThaleMine collects a wide variety of data from public datasets and presents it in a easy-to-interrogate form, facilitating the experiments of both lab-based researchers or bioinformaticians. This tool is build upon the InterMine software framework, which has been widely adopted across other model organisms.

Chris Town and Sergio Contrino provided a hands-on workshop describing the tools on Araport in last year GARNet2016 meeting and their workshop materials can be downloaded here.

Steenackers WJ, Klíma P, Quareshy M, Cesarino I, Kumpf RP, Corneillie S, Araújo P, Viaene T, Goeminne G, Nowack MK, Ljung K, Friml J, Blakeslee JJ, Novák O, Zažímalová E, Napier RM, Boerjan WA, Vanholme B (2016) cis-cinnamic acid is a novel, natural auxin efflux inhibitor that promotes lateral root formation. Plant Physiol. http:/​/​dx.​doi.​org/pp.00943.2016 Open Access
This pan-european collaboration includes members of Richard Napier’s lab at the University of Warwick. They outline the activity of a novel inhibitor of auxin efflux transport called cis-cinnamic acid (c-CA). When c-CA is applied to growth media plants appears to exhibit an auxin-response phenotype yet these experiments show that c-CA is neither an auxin or anti-auxin and in fact blocks local auxin efflux, thus causing buildup of cellular auxin. This effect does not occur with t-CA showing specificity for c-CA and it does not affect long distance auxin transport, which occurs through the phloem. Therefore this paper presents a new pharamolgical tool for the study of in planta auxin transport and homeostasis.

Matthew Terry and Mike Page (University of Southampton) are co-authors on this Japanese-led study that investigates the function of the SUFB subunit of the SUFBCD iron-sulfur cluster. These Fe-S protein clusters play roles in many metabolic processes and the SUFB mutant hmc1 exhibits a defect in chlorophyll biosynthesis due to an accumulation of Mg-containing biosynthetic intermediates. In addition both SUFC- and SUFD-deficient RNAi lines accumulated the same Mg intermediate indicating that the SUFBCD cluster is responsible for this step necessary for chlorophyll production.

Open Access
This paper from is lead by Stefanie Rosa in Caroline Dean’s lab at the John Innes Centre describes a novel method for imaging single molecules of RNA by smFISH. They analyse the localisation of both nascent and mature mRNAs, allowing for analysis of the location of RNA processing and translation.<

This Arabidopsis Research Roundup has five papers that includes two from the John Innes Centre and two from the University of Edinburgh. Firstly Kristen Bomblies’s group at the JIC have investigated the relationship between temperature and meiotic recombination rates. Secondly Veronica Grieneisen and Stan Maree have developed a mathematical model to characterise cell morphologies taken[…]

The first two papers in this weeks Arabidopsis Research Roundup investigate different aspects of the plants response to temperature fluctuations. Firstly Lars Ostergaard (JIC) looks at the influence of temperature in the control of fruit dehiscence whilst Phil Wigge (SLCU) investigates crosstalk between chloroplast and nuclear signaling. The third paper from Ian Henderson (University of[…]

Charles Melnyk discusses a new paper published in PNAS that describes the molecular events that occur during grafting. The paper is entitled ‘Transcriptome dynamics at Arabidopsis graft junctions reveal an intertissue recognition mechanism that activates vascular regeneration‘ http://blog.garnetcommunity.org.uk/wp-content/uploads/2018/03/Melynk_180301.mp3Podcast: Play in new window | DownloadSubscribe: iTunes | Android | RSS

This edition of the Arabidopsis Research roundup beings with a study from SLCU that provides a molecular context to the changes that occur at graft junctions. Second is a study from Edinburgh that reports on the findings of a citizen science plant phenotyping project. Third are two studies from the John Innes Centre that follow-on[…]

This weeks Arabidopsis Research Roundup begins with a study from SLCU that investigates the interaction between nitrate and cytokinin signaling in the shoot meristem. Next is research from Sheffield that studies changes to the macromolecular composition of the photosynthetic apparatus following the transition from dark to light. Third are three papers that include University of[…]

GARNet with support from the Bristol Centre for Agricultural Innovation and New Phytologist are organising a Gene Editing Workshop that will take place at the University of Bristol on March 26th-27th 2018. This workshop is designed to encourage interactions and discussion about the use of CRISPR-Cas9 gene editing in plant systems. We are encouraging ECRs[…]

The cellular mechanics of auxin perception and signaling have been well studied over the past two decades. The pivotal interaction that controls this activity involves the auxin-dependent contact between the TIR1 receptor and a family of transcriptional regulators called AuxIAA proteins. This interaction has been characterised at a structural level with the auxin indole-3-acetic acid[…]

This weeks Arabidopsis Research Roundup begins with two papers from Royal Hollaway University of London that investigate the factors that control leaf development in the dark and the control of PIN1 phosphorylation. Third is a paper from Bristol that demonstrates the translation of research from Arabidopsis into coriander with regard the control of the response[…]

Enrique Lopez-Juez (Royal Holloway University of London) introduces a paper from that attempts to answer a critical question in plant science ‘Why do plants makes leaves in the dark‘? http://www.plantphysiol.org/content/early/2017/12/28/pp.17.01730.long http://blog.garnetcommunity.org.uk/wp-content/uploads/2018/01/Lopez_180116.mp3Podcast: Play in new window | DownloadSubscribe: iTunes | Android | RSS